The immediate goal of this project is to identify new biomarker proteins for type 2 diabetes and early detection of risk of type 2 diabetes and to test a new photoaptamer array marker analysis system. The longer term goal is to foster and validate the measurement of new markers for the risk of type 2 diabetes (and markers found by others) in a robust and reliable photoaptamer array diagnostic platform. The hypothesis underlying this work is that a deep, global search for protein markers will reveal patterns of multiple markers that will signal type 2 diabetes (and other disease) risk states significantly earlier and much more reliably than current diagnostics, without requiring perturbations such as glucose administration and extensive multipoint tolerance measures. Global proteomics methods are well suited for disease marker discovery but appear to be too expensive for direct diagnosis, while the photoaptamer systems appear to be extremely well suited for providing routine, reliable, high-throughput diagnosis. We plan to search more deeply into the blood serum proteome for biomarkers and biomarker isoforms than has been accomplished previously, using new ultrasensitive multiplex fluorescent detection dyes developed at Montana State University-in a 2D gel format with liquid state IEF prefractionation. This improved methodology provides a powerful means to detect differences in expression of proteins between samples and is especially powerful for measuring changes in modified protein isoforms. We will complement and compare the 2D gel analysis with high resolution 2D-LC/MS/MS on the same fractions, using isotopic cysteine and lysine tags for measuring changes in protein and peptide expression. A clinical group at the U Mass Medical Center, Worcester (that is seeking to develop more effective lifestyle modification strategies for people at risk of type 2 diabetes) is providing samples from well characterized patients and controls. This project brings together our academic group that is developing and applying new targeted proteomics detection technologies with outstanding groups of investigators from industry that are contributing advanced technology resources and a powerful diagnostic platform. Relevance to public health: Approximately 5 million people in the US are thought to have undiagnosed Type 2 diabetes and the numbers appear to be growing. At the time of diagnosis of Type 2 diabetes many patients already have severe complications of diabetes and are a greatly increased risk of heart disease. Available screening methods, such as the oral glucose tolerance test appear to be of modest effectiveness, are expensive and difficult to carry out and there is little evidence of benefit from screening asymptomatic adults for undiagnosed diabetes with current technology. This project aims to identify diagostic targets and to foster the development more facile, inexpensive, sensitive, and reliable clinical tests.